OBJECTIVE: To gain information about the components and events needed to activate the expression of viral and cellular eucaryotic genes. APPROACH: This project proposes to separate transcribing chromatin from non-transcribing chromatin by use of an affinity column which binds RNA Polymerase II. SV40 chromosomes as well as fragmented chromatin from tissue culture cells will be used. The biochemical composition as well as structural features of the transcriptionally active and inactive fractions will be characterized. A comparison of this information will be used to attempt to determine what is needed to activate expression of a gene. CURRENT PLANS and PROGRESS: Two types of affinity columns for RNA Polymerase II are being developed. So far one of them has been shown to specifically bind purified RNA Polymerase II, RNA Ploymerase II in a crude extract, and SV 40 viral transcription complexes. Research effort is at present directed toward dealing with non-specific background binding. Future plans are to characterize the biochemical and structural features of the bound and unbound fractions.